Highly Altered Vβ Repertoire of T Cells Infiltrating Long-Term Rejected Kidney Allografts

Abstract

Chronic rejection represents a major cause of long-term kidney graft loss. T cells that are predominant in long-term rejected kidney allografts (35 ± 10% of area infiltrate) may thus be instrumental in this phenomenom, which is likely to be dependant on the indirect pathway of allorecognition only. We have analyzed the variations in T cell repertoire usage of the Vβ chain at the complementary determining region 3 (CDR3) level in 18 human kidney grafts lost due to chronic rejection. We observed a strongly biased intragraft TCR Vβ usage for the majority of Vβ families and also a very high percentage (55%) of Vβ families exhibiting common and oligoclonal Vβ-Cβ rearrangements in the grafts of patients with chronic rejection associated with superimposed histologically acute lesions. Furthermore, Vβ8 and Vβ23 families exhibited common and oligoclonal Vβ-Jβ rearrangements in 4 of 18 patients (22%). Several CDR3 amino acid sequences were found for the common and oligoclonal Vβ8-Jβ1.4 rearrangement. Quantitative PCR showed that biased Vβ transcripts were also overexpressed in chronically rejected kidneys with superimposed acute lesions. In contrast, T lymphocytes infiltrating rejected allografts with chronic rejection only showed an unaltered Gaussian-type CDR3 length distribution. This pattern suggests that late graft failure associated with histological lesions restricted to Banff-defined chronic rejection does not involve T cell-mediated injury. Thus, our observation suggests that a limited number of determinants stimulates the recipient immune system in long-term allograft failure. The possibility of a local response against viral or parenchymatous cell-derived determinants is discussed.

Expansion of Highly Differentiated Cytotoxic Terminally Differentiated Effector Memory CD8+ T Cells in a Subset of Clinically Stable Kidney Transplant Recipients

“…[35][36][37] A capillary sequencer (Applied Biosystems 3730) 38 was used to determine the CDR3 length distribution after amplification of the Vb genes transcript. 39 The level of Vb family transcripts was measured by quantitative RT-PCR and normalized by a reference gene (HPRT).…”

Direct Recognition of Foreign MHC Determinants by Naive T Cells Mobilizes Specific Vβ Families Without Skewing of the Complementarity-Determining Region 3 Length Distribution

“…Analysis of qualitative alterations in the T cell repertoire has made an essential contribution to the understanding of several biological situations including alloimmunity (23), tumor immunity (24), and autoimmune diseases (25). We have previously shown in vivo that T cells infiltrating allografts long after transplantation exhibit a strongly altered TCR V␤ repertoire in both rats and humans (23,26). However, no equivalent data under conditions of direct restricted recognition of foreign APCs are available.…”

“…Aliquots of the cDNA synthesis reaction were amplified with 1 of the 24 human (the pseudo-genes V␤ 10, 20, 21, and 24 were not included in the study) or 1 of the 20 rat V␤ family-specific primers and the corresponding C␤ primer (29,30). The PCR amplification conditions were as previously described (23,26). Immunoscope software (31) was used to obtain a semiquantitative analysis of the V␤ chain of the TCR repertoire at the CDR3.…”

Persistence of Dominant T Cell Clones in Accepted Solid Organ Transplants

“…This common antigenic disparity did not, however, promote dominance of the same V␤ TCR clones in each accepted graft, a possible consequence of the genetic diversity of the TCR loci in these animals. Selective TCR V␤ repertoires have been detected on alloreactive T cells in graft rejection responses (26,27,31). Likewise, preliminary data from our laboratory indicate that clonal dominance occurs among GITC-invading class I disparate kidneys rejected by untreated recipient animals.…”

“…TCR spectratyping has been used in rodents (13, 20 -23) and man (24) to evaluate the TCR␤ chain repertoire usage under normal physiologic conditions (24) and various pathological situations, including graft-vs-host disease (25), allografts (17,26,27), response to viral Ags (28), rheumatoid arthritis (29), and malignancies (18). In the present study, this technology was used to evaluate, in a preclinical model, the TCR V␤ repertoire of intragraft T cells and to assess potential correlates with in situ T cell tolerance.…”